Compositions and methods for inhibition of HAO1 (Hydroxyacid Oxidase 1 (Glycolate Oxidase)) gene expression

Abstract

The invention relates to RNAi agents, e.g. double-stranded RNAi agents, targeting the HAO1 gene, and method of using such RNAi agents to inhibit expression of HAO1 and methods of treating subjects having, e.g., PH1. Described herein are double-stranded RNAi agents which inhibit the expression of a HA01 gene in a cell, such as a cell within a subject, e.g., a mammal, such as a human having a HAO1 associated disorder, and uses of such double-stranded RNAi agents. In certain aspects of the invention, substantially all of the nucleotides of an iRNA of the invention are modified.

We claim: 
     
       1. A double stranded RNAi agent capable of inhibiting expression of HAO1 in a cell, wherein said double stranded RNAi agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand nucleotide sequence consists of SEQ ID NO:589 and the antisense strand nucleotide sequence consists of SEQ ID NO:706;
 wherein substantially all of the nucleotides of said sense strand and substantially all of the nucleotides of said antisense strand are modified nucleotides, and 
 wherein said sense strand is conjugated to a ligand attached at the 3′-terminus. 
 
     
     
       2. The double stranded RNAi agent of  claim 1 , wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand are modified nucleotides. 
     
     
       3. The double stranded RNAi agent of  claim 1 , wherein at least one of said modified nucleotides is selected from the group consisting of a 3′-terminal deoxy-thymine (dT) nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a non-natural base comprising nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a nucleotide comprising a 5′ phosphate or 5′ phosphate mimic, and a terminal nucleotide linked to a cholesteryl derivative or a dodecanoic acid bisdecylamide group. 
     
     
       4. The double stranded RNAi agent of  claim 1 , wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker. 
     
     
       5. The double stranded RNAi agent  claim 1 , wherein the ligand is 
       
         
           
           
               
               
           
         
       
     
     
       6. The double stranded RNAi agent of  claim 1 , wherein the ligand is attached to the 3′ end of the sense strand. 
     
     
       7. The double stranded RNAi agent of  claim 6 , wherein the RNAi agent is conjugated to the ligand as shown in the following schematic 
       
         
           
           
               
               
           
         
         wherein X is O or S. 
       
     
     
       8. The double stranded RNAi agent of  claim 1 , wherein said agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage. 
     
     
       9. The double stranded RNAi agent of  claim 8 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 3′-terminus of one strand. 
     
     
       10. The double stranded RNAi agent of  claim 8 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 5′-terminus of one strand. 
     
     
       11. The double stranded RNAi agent of  claim 8 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the both the 5′- and 3′-terminus of one strand. 
     
     
       12. The double stranded RNAi agent of  claim 1 , wherein the base pair at the 1 position of the 5′-end of the antisense strand of the duplex is an AU base pair. 
     
     
       13. The double stranded RNAi agent of  claim 1 , wherein the sense strand nucleotide sequence comprises SEQ ID NO:589 and the antisense strand nucleotide sequence comprises SEQ ID NO:706. 
     
     
       14. The double stranded RNAi agent of  claim 1 , wherein the sense strand nucleotide sequence consists of SEQ ID NO:589 and the antisense strand nucleotide sequence consists of SEQ ID NO:706. 
     
     
       15. A vector containing the double stranded RNAi agent of  claim 1 . 
     
     
       16. A cell containing the double stranded RNAi agent of  claim 1 . 
     
     
       17. A pharmaceutical composition comprising the double stranded RNAi agent of  claim 1 . 
     
     
       18. A method of inhibiting HAO1 expression in a cell, the method comprising:
 (a) contacting the cell with the double stranded RNAi agent of  claim 1 ; and 
 (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a HAO1 gene, thereby inhibiting expression of the HAO1 gene in the cell. 
 
     
     
       19. The double stranded RNAi agent of  claim 1 , wherein the sense strand consists of SEQ ID NO:213 (gsascuuuCfaUfCfCfuggaaauauaL96) and the antisense strand consists of SEQ ID NO:330 (usAfsuauUfuCfCfaggaUfgAfaagucscsa) and each abbreviation represents a nucleotide monomer as follows: 
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   Abbreviation 
                   Nucleotide(s) 
                 
                     
                     
                 
                     
                   Af 
                   2′-fluoroadenosine-3′-phosphate 
                 
                     
                   Afs 
                   2′-fluoroadenosine-3′-phosphorothioate 
                 
                     
                   Cf 
                   2′-fluorocytidine-3′-phosphate 
                 
                     
                   U 
                   Uridine-3′-phosphate 
                 
                     
                   Uf 
                   2′-fluorouridine-3′-phosphate 
                 
                     
                   a 
                   2′-O-methyladenosine-3′-phosphate 
                 
                     
                   as 
                   2′-O-methyladenosine-3′-phosphorothioate 
                 
                     
                   c 
                   2′-O-methylcytidine-3′-phosphate 
                 
                     
                   cs 
                   2′-O-methylcytidine-3′-phosphorothioate 
                 
                     
                   g 
                   2′-O-methylguanosine-3′-phosphate 
                 
                     
                   gs 
                   2′-O-methylguanosine-3′-phosphorothioate 
                 
                     
                   u 
                   2′-O-methyluridine-3′-phosphate 
                 
                     
                   us 
                   2′-O-methyluridine-3′-phosphorothioate 
                 
                     
                   s 
                   phosphorothioate linkage 
                 
                     
                   L96 
                   N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4- 
                 
                     
                     
                   hydroxyprolinol Hyp-(GalNAc-alkyl)3.